Production of l-lysine and l-lysine-containing feed additives

Abstract

The invention relates to a process for producing L-lysine or L-lysine-containing feed additives by fermentation, which comprises a) expressing a polynucleotide coding for polypeptide having LL-diaminopimelate aminotransferase activity in a bacterium excreting L-lysine, and b) fermenting the resultant bacterium in a medium under suitable conditions and allowing the resultant L-lysine to accumulate in the fermentation broth.

1 - 27 . (canceled) 
     
     
         28 . A process for producing L-lysine or L-lysine-containing feed additives by fermentation, comprising:
 a) expressing a polynucleotide coding for a polypeptide having LL-diaminopimelate aminotransferase activity in an L-lysine excreting bacterium; and   b) fermenting the resultant bacterium in a medium under suitable conditions and allowing the resultant L-lysine excreted by the bacterium to accumulate in the fermentation broth.   
     
     
         29 . The process of  claim 28 , wherein the polynucleotide coding for a polypeptide having LL-diaminopimelate aminotransferase activity is a polynucleotide from  Arabidopsis thaliana, Synechococcus  sp.,  Oryza sativa  or  Vitis vinifera.    
     
     
         30 . The process of  claim 28 , wherein the bacterium is a bacterium selected from the group coryneform bacteria, the family Enterobacteriaceae and the genus  Bacillus.    
     
     
         31 . The process of  claim 30 , wherein the coryneform bacterium is a bacterium of the genus  Corynebacterium.    
     
     
         32 . The process of  claim 31 , wherein the bacterium of the genus  Corynebacterium  is a bacterium selected from the group  Corynebacterium glutamicum, Corynebacterium efficiens, Corynebacterium callunae, Corynebacterium thermoaminogenes  and  Corynebacterium ammoniagenes.    
     
     
         33 . The process of  claim 32 , wherein the bacterium is  Corynebacterium glutamicum.    
     
     
         34 . The process of  claim 30 , wherein the bacterium of the family Enterobacteriaceae is a bacterium of the genus  Escherichia.    
     
     
         35 . The process of  claim 34 , wherein the bacterium of the genus  Escherichia  is  Escherichia coli.    
     
     
         36 . The process of  claim 30 , characterized in that the bacterium of the genus  Bacillus  is  Bacillus subtilis  or  Bacillus methanolicus.    
     
     
         37 . The process of  claim 28 , wherein said bacterium excretes at least 0.25 g/l L lysine in maximally 120 hours. 
     
     
         38 . The process of  claim 28 , wherein bacterium possesses a gene coding for a feedback resistant aspartate kinase. 
     
     
         39 . The process of  claim 38 , wherein the polypeptide has an amino acid sequence which is at least 70% identical to the amino acid sequences selected from the group: SEQ ID NO: 2 SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, and SEQ ID NO: 10. 
     
     
         40 . The process of  claim 39 , wherein the polypeptide has an amino acid sequence which is identical to an amino acid sequence selected from the group: SEQ ID NO: 2 SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, and SEQ ID NO: 10. 
     
     
         41 . The process of  claim 28 , wherein the bacterium is fermented in a fed-batch process. 
     
     
         42 . The process of  claim 28 , wherein the bacterium is fed in a repeated fed-batch process. 
     
     
         43 . The process of  claim 28 , wherein the bacterium is fermented in a continuous process. 
     
     
         44 . The process of  claim 28 , wherein the L-lysine excreted by the bacterium is collected. 
     
     
         45 . The process of  claim 28 , wherein the L-lysine excreted by the bacterium is obtained together with components of the fermentation broth. 
     
     
         46 . The process of  claim 28 , wherein the L-lysine excreted by the bacterium is isolated and purified. 
     
     
         47 . An L-lysine-excreting bacterium which has been transformed by a polynucleotide coding for a polypeptide having LL-diaminopimelate aminotransferase activity and, compared with a non-transformed bacterium, exhibits increased excretion of L-lysine. 
     
     
         48 . The L-lysine-excreting bacterium of  claim 47 , wherein the polypeptide having LL-diaminopimelate aminotransferase activity is a polynucleotide of  Arabidopsis thaliana, Synechococcus  sp.,  Oryza sativa  or  Vitis vinifera.    
     
     
         49 . The L-lysine-excreting bacterium of  claim 47 , wherein the bacterium is a coryneform bacterium. 
     
     
         50 . The L-lysine-excreting bacterium of  claim 49 , wherein the bacterium is a coryneform bacterium of the genus  Corynebacterium.    
     
     
         51 . The L-lysine-excreting bacterium of  claim 50 , wherein the bacterium of the genus  Corynebacterium  is  Corynebacterium glutamicum.    
     
     
         52 . The L-lysine-excreting bacterium of  claim 47 , wherein the bacterium is a bacterium of the family Enterobacteriaceae. 
     
     
         53 . The L-lysine-excreting bacterium of  claim 52 , wherein the bacterium of the family Enterobacteriaceae is of the genus  Escherichia.    
     
     
         54 . The L-lysine-excreting bacterium of  claim 53 , wherein the bacterium of the genus  Escherichia  is  Escherichia coli.